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Find video protocols related to scientific articles indexed in Pubmed.
Self-assembly behavior of a linear-star supramolecular amphiphile based on host-guest complexation.
Langmuir
PUBLISHED: 10-24-2014
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A star polymer, ?-cyclodextrin-poly(l-lactide) (?-CD-PLLA), and a linear polymer, azobenzene-poly(ethylene glycol) (Azo-PEG), could self-assemble into a supramolecular amphiphilic copolymer (?-CD-PLLA@Azo-PEG) based on the host-guest interaction between ?-CD and azobenzene moieties. This linear-star supramolecular amphiphilic copolymer further self-assembled into a variety of morphologies, including sphere-like micelle, carambola-like micelle, naan-like micelle, shuttle-like lamellae, tube-like fiber, and random curled-up lamellae, by tuning the length of hydrophilic or hydrophobic chains. The variation of morphology was closely related to the topological structure and block ratio of the supramolecular amphiphiles. These self-assembly structures could disassemble upon an ultraviolet (UV) light irradiation.
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Highly efficient separation, enrichment, and recovery of peptides by silica-supported polyethylenimine.
Langmuir
PUBLISHED: 10-07-2014
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Highly efficient and charge-selective adsorption and desorption of peptides at trace level by a solid-phase adsorbent is described. The adsorbent of SiO2@PEI is synthesized by covalent immobilization of branched polyethylenimines (PEI) exclusively on the outer surface of the porous silica particles (?300 ?m). For aqueous peptides (Mw = 600-3000 Da), SiO2@PEI can capture the negatively charged ones and leave the positively charged ones intact, and by adjusting pH of the system peptides with different isoelectric points (pIs) can be well separated. Targeted peptide at low abundance (at least as low as 0.1 mol % with respect to the highest one) can be well separated. The association constants of K > 10(12) M(-1) at pH > pI and K < 10(4) M(-1) at pH < pI are found; that is, selectivity > 10(8) is generally available. Thus, a peptide even at sub-femtomolar level can be extracted and eluted for analysis, and efficient recovery (79-92%) of the peptides is found. The extraction is mainly promoted by multisite electrostatic interaction, and the hydrophilic and cationic properties of PEI at low pH play a unique role in desorption efficiency and selectivity. The unbiased nature of this method renders the adsorbent applicable to the efficient separation of a broad spectrum of peptides, including those with similar pIs.
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Bioactive nanoparticle-gelatin composite scaffold with mechanical performance comparable to cancellous bones.
ACS Appl Mater Interfaces
PUBLISHED: 07-29-2014
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Mechanical properties are among the most concerned issues for artificial bone grafting materials. The scaffolds used for bone grafts are either too brittle (glass) or too weak (polymer), and therefore composite scaffolds are naturally expected as the solution. However, despite the intensive studies on composite bone grafting materials, there still lacks a material that could be matched to the natural cancellous bones. In this study, nanosized bioactive particles (BP) with controllable size and good colloidal stability were used to composite with gelatin, forming macroporous scaffolds. It was found that the mechanical properties of obtained composite scaffolds, in terms of elastic modulus, compressive strength, and strain at failure, could match to that of natural cancellous bones. This is ascribed to the good distribution of particle in matrix and strong interaction between particle and gelatin. Furthermore, the incorporation of BPs endues the composite scaffolds with bioactivity, forming HA upon reacting with simulated body fluid (SBF) within days, thus stimulating preosteoblasts attachment, growth, and proliferation in these scaffolds. Together with their good mechanical properties, these composite scaffolds are promising artificial bone grating materials.
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An adaptive failure detector based on quality of service in peer-to-peer networks.
Sensors (Basel)
PUBLISHED: 07-01-2014
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The failure detector is one of the fundamental components that maintain high availability of Peer-to-Peer (P2P) networks. Under different network conditions, the adaptive failure detector based on quality of service (QoS) can achieve the detection time and accuracy required by upper applications with lower detection overhead. In P2P systems, complexity of network and high churn lead to high message loss rate. To reduce the impact on detection accuracy, baseline detection strategy based on retransmission mechanism has been employed widely in many P2P applications; however, Chen's classic adaptive model cannot describe this kind of detection strategy. In order to provide an efficient service of failure detection in P2P systems, this paper establishes a novel QoS evaluation model for the baseline detection strategy. The relationship between the detection period and the QoS is discussed and on this basis, an adaptive failure detector (B-AFD) is proposed, which can meet the quantitative QoS metrics under changing network environment. Meanwhile, it is observed from the experimental analysis that B-AFD achieves better detection accuracy and time with lower detection overhead compared to the traditional baseline strategy and the adaptive detectors based on Chen's model. Moreover, B-AFD has better adaptability to P2P network.
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Spatio-temporal patterns of schistosomiasis japonica in lake and marshland areas in China: the effect of snail habitats.
Am. J. Trop. Med. Hyg.
PUBLISHED: 06-30-2014
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The progress of the integrated control policy for schistosomiasis implemented since 2005 in China, which is aiming at reducing the roles of bovines and humans as infection sources, may be challenged by persistent presence of infected snails in lake and marshland areas. Based on annual parasitologic data for schistosomiasis during 2004-2011 in Xingzi County, a spatio-temporal kriging model was used to investigate the spatio-temporal pattern of schistosomiasis risk. Results showed that environmental factors related to snail habitats can explain the spatio-temporal variation of schistosomiasis. Predictive maps of schistosomiasis risk illustrated that clusters of the disease fluctuated during 2004-2008; there was an extensive outbreak in 2008 and attenuated disease occurrences afterwards. An area with an annually constant cluster of schistosomiasis was identified. Our study suggests that targeting snail habitats located within high-risk areas for schistosomiasis would be an economic and sustainable way of schistosomiasis control in the future.
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Ubiquitous health monitoring and real-time cardiac arrhythmias detection: a case study.
Biomed Mater Eng
PUBLISHED: 06-24-2014
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As the symptoms and signs of heart diseases that cause sudden cardiac death, cardiac arrhythmia has attracted great attention. Due to limitations in time and space, traditional approaches to cardiac arrhythmias detection fail to provide a real-time continuous monitoring and testing service applicable in different environmental conditions. Integrated with the latest technologies in ECG (electrocardiograph) analysis and medical care, the pervasive computing technology makes possible the ubiquitous cardiac care services, and thus brings about new technical challenges, especially in the formation of cardiac care architecture and realization of the real-time automatic ECG detection algorithm dedicated to care devices. In this paper, a ubiquitous cardiac care prototype system is presented with its architecture framework well elaborated. This prototype system has been tested and evaluated in all the clinical-/home-/outdoor-care modes with a satisfactory performance in providing real-time continuous cardiac arrhythmias monitoring service unlimitedly adaptable in time and space.
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Bonding analysis of the donor-acceptor sandwiches CpE-MCp (E = B, Al, Ga; M = Li, Na, K; Cp = ??-C?H?).
J Mol Model
PUBLISHED: 06-07-2014
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The nature of E-M bonds in CpE-MCp (E = B, Al, Ga; M = Li, Na, K; Cp = ? (5)-C5H5) donor-acceptor sandwiches was studied using the atoms in molecules (AIM) theory, electron localization function (ELF), energy decomposition analysis (EDA), and natural bond orbital analysis (NBO) methods. Both topological and orbital analysis show that the E atom determines the bond strength of the E-M bonds, while the M atom has little influence on it. E-M bond strength decreases in the order E = B, Al, and Ga. The EDA analysis shows that the electrostatic character decreases following the sequence E = B > Al > Ga. Not only the s orbital, but also the p orbital of the E/M atom participates in formation of the E-M bond. The interactions of E and M with Cp are different. The M-Cp interaction is purely electrostatic while the E-Cp interaction has a partly covalent character.
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Anti-lymphoproliferative activity of alpha-2-macroglobulin in the plasma of hibernating 13-lined ground squirrels and woodchucks.
Vet. Immunol. Immunopathol.
PUBLISHED: 05-20-2014
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Plasma from hibernating (HIB) woodchucks (Marmota monax) or 13-lined ground squirrels (Ictidomys tridecemlineatus) suppressed (3)H-thymidine uptake in mouse spleen cell cultures stimulated with Concanavalin A (ConA); plasma from non-hibernating animals were only slightly inhibitory. Maximum inhibition occurred when HIB plasma was added to the cultures prior to ConA. After HPLC size exclusion chromatography of the HIB ground squirrel plasma, a single fraction (fraction-14) demonstrated inhibitory activity. Assay of fraction-14 from 8 HIB squirrels showed inhibition ranging from 13 to 95%; inhibition was correlated to the time the squirrels were exposed to cold prior to hibernation. Western blot analysis showed the factor to be a large molecular weight protein (>300 kDa), and mass spectrometry identified sequences that were 100% homologous with alpha-2-macroglobulin from humans and other species. These findings indicate a hibernation-related protein that may be responsible for immune system down regulation.
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POSS dendrimers constructed from a 1 ? 7 branching monomer.
Chem. Commun. (Camb.)
PUBLISHED: 04-30-2014
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Here, we demonstrate the first known approach to create G1 and G2 POSS dendrimers with 9 and 65 POSS units, and 56 and 392 terminal vinyl groups from a 1 ? 7 branching monomer in only one and three steps.
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BH3-only molecule Bim mediates ?-cell death in IRS2 deficiency.
Diabetes
PUBLISHED: 04-23-2014
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Irs2-deficient mice develop type 2-like diabetes due to a reduction in ?-cell mass and a failure of pancreatic islets to undergo compensatory hyperplasia in response to insulin resistance. In order to define the molecular mechanisms, we knocked down Irs2 gene expression in mouse MIN6 insulinoma cells. Insulin receptor substrate 2 (IRS2) suppression induced apoptotic cell death, which was associated with an increase in expression of the BH3-only molecule Bim. Knockdown (KD) of Bim reduced apoptotic ?-cell death induced by IRS2 suppression. In Irs2-deficient mice, Bim ablation restored ?-cell mass, decreased the number of TUNEL-positive cells, and restored normal glucose tolerance after glucose challenge. FoxO1 mediates Bim upregulation induced by IRS2 suppression, and FoxO1 KD partially inhibits ?-cell death induced by IRS2 suppression. These results suggest that Bim plays an important role in mediating the increase in ?-cell apoptosis and the reduction in ?-cell mass that occurs in IRS2-deficient diabetes.
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Synthesis of Indolyldiketopiperazines with NBS.
Chirality
PUBLISHED: 04-22-2014
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Two series of indolyldiketopiperazines were synthesized starting from methyl 1-substituted-1,2,3,4-tetrahydro-?-carboline-3-carboxylate hydrochlorides via N-bromo-succinimide (NBS) as an important reagent. All eight compounds were characterized by nuclear magnetic resonance (NMR) and elemental analysis. Furthermore, the mechanisms of NBS-reacted rearrangements are also discussed. Chirality 00:000-000, 2014. © 2014 Wiley Periodicals, Inc.
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Modelling and laboratory studies on the adhesion fatigue performance for thin-film asphalt and aggregate system.
ScientificWorldJournal
PUBLISHED: 04-18-2014
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Adhesion between asphalt and aggregate plays an important role in the performance of asphalt mixtures. A low-frequency adhesion fatigue test was proposed in this paper to study the effect of environment on the asphalt-aggregate adhesion system. The stress-based fatigue model had been utilized to describe the fatigue behavior of thin-film asphalt and aggregate system. The factors influencing the adhesion fatigue performance were also investigated. Experiment results show that asphalt has more important effect on the adhesion performance comparing with aggregate. Basalt, which is regarded as hydrophobic aggregates with low silica content, has better adhesion performance to asphalt binder when compared with granite. The effects of aging on the adhesion fatigue performance are different for PG64-22 and rubber asphalt. Long-term aging is found to reduce the adhesion fatigue lives for rubber asphalt and aggregate system, while the effect of long-term aging for aggregate and PG64-22 binder system is positive. Generally the increased stress amplitude and test temperature could induce greater damage and lead to less fatigue lives for adhesion test system.
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Spatiotemporal trends of terrestrial vegetation activity along the urban development intensity gradient in China's 32 major cities.
Sci. Total Environ.
PUBLISHED: 04-15-2014
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Terrestrial vegetation plays many pivotal roles in urban systems. However, the impacts of urbanization on vegetation are poorly understood. Here we examined the spatiotemporal trends of the vegetation activity measured by MODIS Enhanced Vegetation Index (EVI) along Urban Development Intensity (UDI) gradient in 32 major Chinese cities from 2000 to 2012. We also proposed to use a new set of concepts (i.e., actual, theoretical, and positive urbanization effects) to better understand and quantify the impacts of urbanization on vegetation activities. Results showed that the EVI decreased significantly along a rising UDI for 28 of 32 cities (p<0.05) in linear, convex or concave form, signifying the urbanization impacts on vegetation varied across cities and UDI zones within a city. Further, the actual urbanization effects were much weaker than the theoretical estimates because of the offsetting positive effects generated by multiple urban environmental and anthropogenic factors. Examining the relative changes of EVI in various UDI zones against that in the rural area (?EVI), which effectively removed the effects of climate variability, demonstrated that ?EVI decreased markedly from 2000 to 2012 for about three-quarters of the cities in the exurban (0.05
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Gelatin functionalized graphene oxide for mineralization of hydroxyapatite: biomimetic and in vitro evaluation.
Nanoscale
PUBLISHED: 04-05-2014
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We report a facile modification of graphene oxide (GO) by gelatin to mimic charged proteins present in the extracellular matrix during bone formation. The bioinspired surface of GO-gelatin (GO-Gel) composite was used for biomimetic mineralization of hydroxyapatite (HA). A detailed structural and morphological characterization of the mineralized composite was performed. Additionally, MC3T3-E1 cells were cultured on the GO-Gel surfaces to observe various cellular activities and HA mineralization. Higher cellular activities such as cell adhesion, cell proliferation, and alkaline phosphatase activity (ALP) were observed on the GO-Gel surface compared with the GO or glass surface. The increase of ALP confirms that the proposed GO-Gel promotes the osteogenic differentiation of MC3T3-E1 cells. Moreover, the evidence of mineralization evaluated by scanning electron microscopy (SEM) and alizarin red staining (ARS) corroborate the idea that a native osteoid matrix is ultimately deposited. All these data suggest that the GO-Gel hybrids will have great potential as osteogenesis promoting scaffolds for successful application in bone surgery.
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Coxsackievirus-induced miR-21 disrupts cardiomyocyte interactions via the downregulation of intercalated disk components.
PLoS Pathog.
PUBLISHED: 04-01-2014
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Intercalated disks (ICDs) are substantial connections maintaining cardiac structures and mediating signal communications among cardiomyocytes. Deficiency in ICD components such as desmosomes, fascia adherens and gap junctions leads to heart dysfunction. Coxsackievirus B3 (CVB3) infection induces cardiac failure but its pathogenic effect on ICDs is unclear. Here we show that CVB3-induced miR-21 expression affects ICD structure, i.e., upregulated miR-21 targets YOD1, a deubiquitinating enzyme, to enhance the K48-linked ubiquitination and degradation of desmin, resulting in disruption of desmosomes. Inhibition of miR-21 preserves desmin during CVB3 infection. Treatment with proteasome inhibitors blocks miR-21-mediated desmin degradation. Transfection of miR-21 or knockdown of YOD1 triggers co-localization of desmin with proteasomes. We also identified K108 and K406 as important sites for desmin ubiquintination and degradation. In addition, miR-21 directly targets vinculin, leading to disturbed fascia adherens evidenced by the suppression and disorientation of pan-cadherin and ?-E-catenin proteins, two fascia adherens-components. Our findings suggest a new mechanism of miR-21 in modulating cell-cell interactions of cardiomyocytes during CVB3 infection.
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Bioactive nanoparticle through postmodification of colloidal silica.
ACS Appl Mater Interfaces
PUBLISHED: 03-27-2014
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Bioactive nanoparticles with controllable size and good colloidal stability were synthesized through surface modification of colloidal silica nanoparticles with Ca(OH)2 as the modifier. These modified nanoparticles showed good bioactivity, showing evidence of hydroxyapatite formation when incubated in simulated body fluid within 3 days. Comparison of bioactivity was made among different sized particles from nanoscale to microscale. It was found the bioactivity of these calcium modified colloidal silica particles generally decreased with particle size in the explored size range (40 nm particles showed bioactivity within 1 day). These particles were also found to be noncytotoxic but promote preosteoblast growth, thus making them promising bioactive additives for bone repair materials.
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The fruits of wampee inhibit H2O2-induced apoptosis in PC12 cells via the NF-?B pathway and regulation of cellular redox status.
Molecules
PUBLISHED: 03-24-2014
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Wampee (Clausena lansium) fruits (CLS), whose pulp can be used to prepare fruit cups, desserts, jam, or jelly, can be eaten along with the peel. In this study, a PC12 cell model was built to observe the protective effect of CLS against H2O2-induced oxidative stress. We found that pretreatment with CLS increased cell viability and inhibited cytotoxicity, caspase-3 activity and DNA condensation. CLS also attenuated the increase in ROS production and MMP reduction. Moreover, we attempted to determine whether CLS suppressed the expression and phosphorylation of NF-?B. Western blot and immunostaining assay revealed that CLS inhibited H2O2-induced up-regulation of NF-?B p65 and pNF-?B p65. And CLS significantly suppressed the translocation of NF-?B p65 and pNF-?B p65 from cytoplasm to nuclear. Also, seven major compounds including a new flavanoid, luteolin-4'-O-?-d-gluco-pyranoside (3) and six known compounds 1,2, 4-7 were isolated and identified from CLS. Their antioxidative and H2O2-induced PC12 cell apoptosis-reversing activity were determined. These findings suggest that CLS and its major constituents (flavanoids) may be potential antioxidant agents and should encourage further research into their use as a functional food for neurodegenerative diseases.
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Role of BH3-only molecules Bim and Puma in ?-cell death in Pdx1 deficiency.
Diabetes
PUBLISHED: 03-21-2014
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Mutations in pancreatic duodenal homeobox-1 (PDX1) are associated with diabetes in humans. Pdx1-haploinsufficient mice develop diabetes due to an increase in ?-cell death leading to reduced ?-cell mass. For definition of the molecular link between Pdx1 deficiency and ?-cell death, Pdx1-haploinsufficient mice in which the genes for the BH3-only molecules Bim and Puma had been ablated were studied on a high-fat diet. Compared with Pdx1(+/-) mice, animals haploinsufficient for both Pdx1 and Bim or Puma genes showed improved glucose tolerance, enhanced ?-cell mass, and reduction in the number of TUNEL-positive cells in islets. These results suggest that Bim and Puma ablation improves ?-cell survival in Pdx1(+/-) mice. For exploration of the mechanisms responsible for these findings, Pdx1 gene expression was knocked down in mouse MIN6 insulinoma cells resulting in apoptotic cell death that was found to be associated with increased expression of BH3-only molecules Bim and Puma. If the upregulation of Bim and Puma that occurs during Pdx1 suppression was prevented, apoptotic ?-cell death was reduced in vitro. These results suggest that Bim and Puma play an important role in ?-cell apoptosis in Pdx1-deficient diabetes.
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Enhancing effects of hydrogen/halogen bonds on ?-hole interactions involving ylide.
J Mol Model
PUBLISHED: 03-13-2014
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Cooperativity between the H/Cl···C bonds of XH/XCl···CH2PH3 and the P···N interaction of CH2PH3···NH3 in XH/XCl···CH2PH3···NH3 (X = F, N3, CN, CCCN, CCF) was investigated by performing second-order Møller-Plesset perturbation theory (MP2) calculations and quantum theory of atoms in molecules (QTAIM) studies. The formation of a hydrogen/halogen bond greatly extends the scope and increases the most positive electrostatic potential of the ?-hole on the outer surface of the phosphorus atom. This increases the P···N interaction energy, the electron density at the P···N bond critical point, the electrostatic character of the P···N interaction, and it decreases the P···N interaction distance. The net result is that the formation of a hydrogen/halogen bond enhances the P···N interaction, and vice versa. However, the P···N interaction is enhanced by the presence of the hydrogen or halogen bond to a much greater degree than the hydrogen or halogen bond is enhanced by the P···N interaction.
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Single amino acid substitution of VP1 N17D or VP2 H145Y confers acid-resistant phenotype of type Asia1 foot-and-mouth disease virus.
Virol Sin
PUBLISHED: 03-12-2014
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Infection by foot-and-mouth disease virus (FMDV) is triggered by the acidic pH in endosomes after virus uptake by receptor-mediated endocytosis. However, dissociation of the FMDV 146S particle in mildly acidic conditions renders inactivated foot-and-mouth disease (FMD) vaccines much less effective. Type Asia1 FMDV mutants with increased resistance to acid inactivation were selected to study the molecular basis of viral resistance to acid-induced disassembly and improve the acid stability of FMDV. Sequencing of capsid-coding regions revealed four amino acid replacements (VP1 N17D, VP2 H145Y, VP2 G192D, and VP3 K153E) in the viral population of the acid-selected 10th passage. We performed single or combined mutagenesis using a reverse genetic system, and our results provide direct experimental evidence that VP2 H145Y or VP1 N17D substitution confers an acid-resistant phenotype to type Asia1 FMDV.
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Unsaturated guluronate oligosaccharide enhances the antibacterial activities of macrophages.
FASEB J.
PUBLISHED: 03-05-2014
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Alginate from marine seaweeds is receiving continuous attention owing to its wide physiological activities. Herein, we sought to elucidate possible effects of alginate-derived polyguluronate (PG) and unsaturated guluronate oligosaccharide (GOS) on antibacterial activities of macrophages. Our results showed that, in contrast to PG, GOS markedly increased the phagocytosis of IgG-opsonized Escherichia coli and Staphylococcus aureus and further inhibited the survival of intracellular bacteria in macrophages. In line with this, GOS treatment resulted in the enhanced expression of Fc? receptors on macrophages. In addition, GOS activated NF-?B pathway, induced TNF-? secretion, and elevated the expression of inducible nitric oxide synthase and the production of nitric oxide. Meanwhile, GOS stimulated the production of reactive oxygen species in macrophages. Moreover, guluronate trimer to hexamer (G3-G6) in GOS exhibited significant activity that increased the bacterial phagocytosis of macrophages, with the pentamer (G5), displaying the highest activity. Finally, our in vivo results further confirmed that GOS but not PG significantly improved bacterial clearance in murine acute peritonitis. In conclusion, GOS enhances antibacterial activities of macrophages via modulating signaling pathways related to innate immunity, suggesting that GOS might be a promising therapeutic candidate to improve the host defense against bacterial infection.
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Foot-and-mouth disease virus low-fidelity polymerase mutants are attenuated.
Arch. Virol.
PUBLISHED: 02-26-2014
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Previous studies have shown that RNA viruses can be attenuated by either increased or decreased viral polymerase replication fidelity. Although foot-and-mouth disease virus (FMDV) high-fidelity RNA-dependent RNA polymerase (RdRp) variants with an attenuated phenotype have been isolated using mutagens, no FMDV mutant with a low-fidelity polymerase has been documented to date. Here, we describe the generation of several FMDV RdRp mutants using site-directed mutagenesis via a reverse genetic system. Mutation frequency assays confirmed that five rescued FMDV RdRp mutant populations had lower replication fidelity than the wild-type virus population, which allowed us to assess the effects of the change in replication fidelity on the virus phenotype. These low-fidelity FMDV RdRp mutants showed increased sensitivity to ribavirin or 5-fluorouracil (5-FU) treatment without a loss of growth capacity in cell cultures. In addition, decreased fitness and attenuated virulence were observed for the RdRp mutants with lower fidelity. Importantly, based on a quantitative analysis for fidelity and virulence, we concluded that lower replication fidelity is associated with a more attenuated virus phenotype. These results further contribute to our understanding of the replication fidelity of polymerases of RNA viruses and its relationship to virulence attenuation.
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Biomimetic and cell-mediated mineralization of hydroxyapatite by carrageenan functionalized graphene oxide.
ACS Appl Mater Interfaces
PUBLISHED: 02-24-2014
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In bone tissue engineering, it is imperative to design multifunctional biomaterials that can induce and assemble bonelike apatite that is close to natural bone. In this study, graphene oxide (GO) was functionalized by carrageenan. The resulting GO-carrageenan (GO-Car) composite was further used as a substrate for biomimetic and cell-mediated mineralization of hydroxyapatite (HA). It was confirmed that carrageenan on the GO surface facilitated the nucleation of HA. The observation of the effect of the GO-Car on the adhesion, morphology, and proliferation of MC3T3-E1 cells was investigated. In vitro studies clearly show the effectiveness of GO-Car in promoting HA mineralization and cell differentiation. The results of this study suggested that the GO-Car hybrid will be a promising material for bone regeneration and implantation.
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Structural determinants of human ?-globin mRNA stability.
J Hematol Oncol
PUBLISHED: 02-20-2014
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The normal accumulation of adult ? and ? globins in definitive erythrocytes is critically dependent upon processes that ensure that the cognate mRNAs are maintained at high levels in transcriptionally silent, but translationally active progenitor cells. The impact of these post-transcriptional regulatory events on the expression of embryonic ? globin is not known, as its encoding mRNA is not normally transcribed during adult erythropoiesis. Recently, though, ? globin has been recognized as a potential therapeutic for ? thalassemia and sickle-cell disease, raising practical questions about constitutive post-transcriptional processes that may enhance, or possibly prohibit, the expression of exogenous or derepresssed endogenous ?-globin genes in definitive erythroid progenitors.
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Antiviral activity of an isatin derivative via induction of PERK-Nrf2-mediated suppression of cap-independent translation.
ACS Chem. Biol.
PUBLISHED: 02-18-2014
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We report here an isatin derivative 45 (ID45) against coxsackievirus B3 (CVB3) replication, which was synthesized based on a high-throughput screen of a unique natural product library. ID45 showed the most potent anti-CVB3 activity among the four synthesized compounds. Treatment of cells with ID45 before or after infection significantly reduced viral particle formation, resulting in protection of cells from virus-induced apoptosis. In addition, ID45 treatment caused remarkable up-regulation of glucose-regulated protein 78 (GRP78), a hallmark of endoplasmic reticulum (ER) stress and an indicator of enhanced cell viability. In identifying the ER stress response pathway induced by ID45, we found that ID45 activated PKR-like ER protein kinase (PERK) but failed to up-regulate eIF2? phosphorylation. Instead ID45 activated transcription factor Nrf2 (NF-E2-related factor-2), which is evidenced by its nuclear translocation and upregulation of its downstream target genes NQO1 (NAD(P)H quinone-oxidoreductase 1) and GCLM (glutamate-cysteine ligase, modifier subunit). This observation was further verified by using siRNAs of GRP78 or Nrf2, which blocked both the translocation of Nrf2 and up-regulation of its target genes, leading to aggressive viral replication and enhanced cell apoptosis. Finally, we found that ID45-induced up-regulation of NQO1 protected eIF4GI, a eukaryotic cap-dependent translation initiation factor, from cleavage by CVB3 protease and degradation by proteasomes. Taken together, our findings established that a novel antiviral mechanism of isatin derivative ID45 inhibits CVB3 replication by promoting cell survival through a PERK/Nrf2-dependent ER stress pathway, which benefits host cap-dependent translation but suppresses CVB3 cap-independent translation.
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A coumarin-derived fluorescent chemosensor for selectively detecting Cu²?: synthesis, DFT calculations and cell imaging applications.
Talanta
PUBLISHED: 02-17-2014
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A novel coumarin-based fluorescent probe L ((4E)-4-((7-hydroxy-4-methyl-2-oxo-2H-chromen-8-yl) methyleneamino)-1,2-dihyydro-2,3-dimethyl-1-phenylpyrazol-5-one) has been developed as a simple and efficient chemosensor which exhibits a significant fluorescence reduction in the presence of metal cations. This sensor exhibits high selectivity and sensitivity toward Cu(2+) over other common cations. The mechanism for detecting copper was evaluated by time-dependent density functional theory (TD-DFT) calculations and the coordination mode was also confirmed by density functional theory (DFT) calculations. Furthermore, results of cell imaging in this study indicate that this new probe may be useful for detection and monitoring of Cu(2+) in biological applications.
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Study of effect of salvianolic acid B on motor function recovery in rats with spinal cord injury.
Biomed Res Int
PUBLISHED: 02-06-2014
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In this study effect of salvianolic acid B was observed on motor function recovery of rats with spinal cord injury. 50 rats were selected and after inducing SCI their recovery under controlled conditions was studied using Sal B and PBS (as control). Both compounds were introduced intraperitoneally in respective groups of traumatic rats at the same time intervals for 28 days. It was observed that Sal B introduced at 5??mg/kg/day resulted in better motor function recovery. BBB score was recorded which increased significantly along with the reduction in cavity area observed by bright field microscopy of tissues, that is, from 1 to 10 and from 0.20 ± 0.05?mm(2) to 0.10 ± 0.03?mm(2), in Sal B treated group, respectively, compared to PBS group. Statistical analysis was carried out using SPSS software (SPSS, Chicago, IL, USA), values were expressed as mean ± SEM, and P value <0.01 was considered significant. Effect of Sal B on expression of NF-kB p65 and IkB ? was studied and OD values of densitometry of western blots were taken. MPO activity was also studied. It was observed that treatment of Sal B significantly reduced the expression of both compounds in Sal B treated group as compared to control group after 28 days of treatment.
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Posterior short segment pedicle screw fixation and TLIF for the treatment of unstable thoracolumbar/lumbar fracture.
BMC Musculoskelet Disord
PUBLISHED: 02-06-2014
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Currently, Posterior Short Segment Pedicle Screw Fixation is a popular procedure for treating unstable thoracolumbar/lumbar burst fracture. But progressive kyphosis and a high rate of hardware failure because of lack of the anterior column support remains a concern. The efficacy of different methods remains debatable and each technique has its advantages and disadvantages.
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Simultaneous determination and pharmacokinetic study of three isoflavones from Trifolium pratense extract in rat plasma by LC-MS/MS.
Biomed. Chromatogr.
PUBLISHED: 01-31-2014
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A highly selective and sensitive liquid chromatography-tandem mass spectrometry has been developed and validated for simultaneous determination of three isoflavones - ononin, formononetin and biochanin A - in rat plasma using lysionotin as internal standard (IS). The plasma samples were pretreated and extracted by liquid-liquid extraction. Chromatographic separation was accomplished on a C18 column with the column temperature of 30?°C and a mobile phase of methanol-0.1% formic acid (75:25, v/v). The detection was accomplished by multiple-reaction monitoring scanning with positive/negative ion-switching electrospray ionization mode. The optimized mass transition ion pairs (m/z) for quantitation were 431.3/269.1 for ononin, 267.1/252.2 for formononetin, 283.2/268.2 for biochanin A and 343.2/313.3 for IS. The total run time was 8.0?min. Full validation of the assay was implemented, including selectivity, sensitivity, linearity, precision, accuracy, recovery, matrix effect and stability. This is the first report on simultaneous determination of the three major isoflavones in rat plasma after intragastric administration of Trifolium pratense extract. The results provided a significant basis for the clinical application of this herb Trifolium pratense. Copyright © 2014 John Wiley & Sons, Ltd.
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Ribavirin-resistant variants of foot-and-mouth disease virus: the effect of restricted quasispecies diversity on viral virulence.
J. Virol.
PUBLISHED: 01-22-2014
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Mutagenic nucleoside analogues can be used to isolate RNA virus high-fidelity RNA-dependent RNA polymerase (RdRp) variants, the majority of which are attenuated in vivo. However, attenuated foot-and-mouth disease virus (FMDV) high-fidelity RdRp variants have not been isolated, and the correlations between RdRp fidelity and virulence remain unclear. Here, the mutagen ribavirin was used to select a ribavirin-resistant population of FMDV, and 4 amino acid substitutions (D5N, A38V, M194I, and M296V) were identified in the RdRp-coding region of the population. Through single or combined mutagenesis using a reverse genetics system, we generated direct experimental evidence that the rescued D5N, A38V, and DAMM mutants but not the M194I and M296V mutants are high-fidelity RdRp variants. Mutagen resistance assays revealed that the higher replication fidelity was associated with higher-level resistance to ribavirin. In addition, significantly attenuated fitness and virulence phenotypes were observed for the D5N, A38V, and DAMM mutants. Based on a systematic quantitative analysis of fidelity and virulence, we concluded that higher replication fidelity is associated with a more attenuated virus. These data suggest that the resulting restricted quasispecies diversity compromises the adaptability and virulence of an RNA virus population. The modulation of replication fidelity to attenuate virulence may represent a general strategy for the rational design of new types of live, attenuated vaccine strains.
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A novel approach for enriching cancer stem cells from the human SW-13 adrenocortical carcinoma cell line.
Anticancer Res.
PUBLISHED: 01-10-2014
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The present study was undertaken to develop a new method for enriching cancer stem cells (CSCs) from the human adrenal cortical carcinoma (ACC) cell line SW-13. Given that the existence of CSCs in ACC causes resistance to conventional chemotherapies, treatment with cyclophosphamide was used for in vivo selection of CSCs in a BALB/c nude mouse tumor xenograft model established using the ACC cell line SW-13. The characteristics of CSCs in three generations of tumor xenografts were assessed for single-cell colony formation, flat colony formation, and cell sphere formation in serum-free suspension culture. The formation rates of single-cell colonies, flat colonies, and cell spheres were significantly higher for tumor xenograft cells treated with cyclophosphamide than for untreated engrafted tumor cells. Flow cytometry to examine expression of the CSC markers C-X-C chemokine receptor type-4 (CXCR4; CD184) and ATP-binding cassette sub-family G member-2 (ABCG2; CDw338) revealed markedly higher levels of CXCR4 and ABCG2 in cyclophosphamide-treated xenograft tumor cells compared to untreated tumor cells. Together, these results indicate that cyclophosphamide treatment of tumor xenograft cells caused enrichment of CSCs with a strong capability for self-renewal and proliferation. In this method, the administration of cyclophosphamide selectively kills cancer cells without toxicity to CSCs and thereby provides a practical approach for achieving the enrichment of CSCs in ACC.
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EFFECTS OF FREQUENCY, IRRADIATION GEOMETRY AND POLARISATION ON COMPUTATION OF SAR IN HUMAN BRAIN.
Radiat Prot Dosimetry
PUBLISHED: 01-09-2014
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The power absorbed by the human brain has possible implications in the study of the central nervous system-related biological effects of electromagnetic fields. In order to determine the specific absorption rate (SAR) of radio frequency (RF) waves in the human brain, and to investigate the effects of geometry and polarisation on SAR value, the finite-difference time-domain method was applied for the SAR computation. An anatomically realistic model scaled to a height of 1.70 m and a mass of 63 kg was selected, which included 14 million voxels segmented into 39 tissue types. The results suggested that high SAR values were found in the brain, i.e. ?250 MHz for vertical polarisation and 900-1200 MHz both for vertical and horizontal polarisation, which may be the result of head resonance at these frequencies.
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A highly selective fluorescence "turn-on" probe for Cu(II) based on reaction and its imaging in living cells.
Inorg Chem
PUBLISHED: 10-11-2013
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A new oxidative C-O bond cleavage reaction-based probe FluHMPP was designed and prepared. FluHMPP displays excellent selective turn-on fluorescence response for Cu(II) in aqueous solution under visible light excitation. The cleavage products are fully characterized. Fluorescein fragment is further oxidized to highly fluorescent MFME (3-O-methylfluorescein methyl ester), and benzyl ether of imine fragment has been transformed to carboxyl. Confocal microscopy experiments have demonstrated that FluHMPP could also be used in live cells for the detection of Cu(II).
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Detection of intestinal intraepithelial lymphocytes, goblet cells and secretory IgA in the intestinal mucosa during Newcastle disease virus infection.
Avian Pathol.
PUBLISHED: 10-02-2013
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Newcastle disease, which is caused by Newcastle disease virus (NDV), is a highly contagious viral disease of poultry and other bird species. The mucosa is the first line of defence to invading pathogens, including NDV, and it has been confirmed that the mucosa can contribute to host protection. This study was conducted to evaluate the intestinal mucosal immunology in NDV infection. Forty specific-pathogen-free chickens were divided into two groups, 20 birds in each group. Group 1 was inoculated with NDV by the intravenous route. Group 2 was used as the control group and was given sterile phosphate-buffered saline by the same route. At 24, 48, 72, and 96 h post infection (h.p.i.), five chickens from each treatment were killed. Samples of the duodenum, jejunum, and ileum were collected to quantify intestinal intraepithelial lymphocytes (IEL), goblet cells and secretory IgA (sIgA) by cytochemistry and immunohistochemistry analysis. The results indicated that IEL were increased from 24 to 72 h.p.i. in the infected tissues, and were significantly higher than in the control group at 48 h.p.i. (P < 0.01). In contrast to IEL, goblet cell numbers were reduced dramatically from 24 to 96 h.p.i. in the infected birds (P < 0.01) Furthermore, the content of sIgA was significantly higher at 48 and 72 h.p.i. in the infected tissues (P < 0.01). sIgA positivity was observed in the epithelial lining of the intestinal mucosa. These data suggest that IEL, goblet cells, and sIgA were involved in the intestinal mucosal immunity against NDV infection.
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VP1 B-C and D-E loops of bovine enterovirus cluster B can effectively display foot-and-mouth disease virus type O-conserved neutralizing epitope.
J. Gen. Virol.
PUBLISHED: 09-28-2013
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On the basis of generation of an infectious cDNA clone for the BHM26 strain of bovine enterovirus cluster B (BEV-B), 22 sites on different loops of the BHM26 capsid were selected according to an alignment of its sequence with the structural motifs of BEV-A strain VG-5-27 for insertion of the foot-and-mouth disease virus (FMDV) type O-conserved neutralizing epitope 8E8. Two recombinant viruses, rBEV-A1 and rBEV-DE, in which the FMDV epitope was inserted into the VP1 B-C or D-E loops, were rescued by transfection of BHK-21 cells with the in vitro-transcribed RNA of the recombinant BHM26 genome-length cDNA constructs. The two epitope-inserted viruses were genetically stable and exhibited growth properties similar to those of their parental virus in BHK-21 and IBRS-2 cells, which are susceptible to both BEV and FMDV. However, the two recombinant BEVs (rBEVs) had a significantly lower growth titre than those of the parental virus BHM26 in MDBK and Marc145 cells, which are susceptible to BEV but not to FMDV. These results indicated that insertion of the FMDV epitope into the VP1 B-C or D-E loops of the BEV particle altered the replication properties of BEV. In addition, the two rBEVs were sensitive to neutralization by the FMDV type O-specific mAb 8E8, and anti-FMDV IgG antibodies were induced in mice by intramuscular inoculation with the rBEV-A1 and rBEV-DE viruses. Our results demonstrate that the VP1 B-C and D-E loops of the BEV-B particle can effectively display a foreign epitope, making this an attractive approach for the design of BEV-vectored and epitope-based vaccines.
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Design of D-?-A type photoacid generators for high efficiency excitation at 405 nm and 800 nm.
Chem. Commun. (Camb.)
PUBLISHED: 08-14-2013
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New sulfonium salts with diphenylamino asymmetrically substituted stilbene as a D-?-A conjugated system have been synthesized. The resulting photoacid generators exhibit a highly efficient acid photogeneration process by either one-photon 405 nm or two-photon 800 nm excitation.
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Forest cutting and impacts on carbon in the eastern United States.
Sci Rep
PUBLISHED: 08-12-2013
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Forest cutting is a major anthropogenic disturbance that affects forest carbon (C) storage and fluxes. Yet its characteristics and impacts on C cycling are poorly understood over large areas. Using recent annualized forest inventory data, we estimated cutting-related loss of live biomass in the eastern United States was 168?Tg C yr(-1) from 2002 to 2010 (with C loss per unit forest area of 1.07?Mg ha(-1) yr(-1)), which is equivalent to 70% of the total U.S. forest C sink or 11% of the national annual CO2 emissions from fossil-fuel combustion over the same period. We further revealed that specific cutting-related C loss varied with cutting intensities, forest types, stand ages, and geographic locations. Our results provide new insights to the characteristics of forest harvesting activities in the eastern United States and highlight the significance of partial cutting to regional and national carbon budgets.
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Peptide hydrogels assembled from nonionic alkyl-polypeptide amphiphiles prepared by ring-opening polymerization.
Biomacromolecules
PUBLISHED: 07-08-2013
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Three alkyl-polypeptide (AP) amphiphiles were prepared using ring-opening polymerization of ?-amino acid N-carboxyanhydride. The polypeptide segment was composed of diethylene-glycol-monomethyl-ether-functionalized poly-L-glutamate (poly-L-EG(2)Glu). These AP amphiphiles can spontaneously self-assemble into transparent hydrogels in water. These hydrogels showed shear thinning properties, and their strength can be modulated by hydrophobic alkyl tails. CryoTEM and AFM characterizations suggested that these hydrogels were formed by nanoribbons arising from intermolecular interactions between nonionic poly-L-EG(2)Glu segments.
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Canal preparation for prosthetic radial head replacement: rasping versus reaming.
J Shoulder Elbow Surg
PUBLISHED: 06-12-2013
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While many design-specific features of radial head prostheses have been studied (ie, geometry and surface coating), the optimum technique for canal preparation has not been determined. We hypothesized that preparation of the radial canal with a reamer would allow for the accommodation of a larger stem diameter versus following canal preparation with a rasp, and would provide acceptable stem-bone micromotion.
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Redox-responsive hyperbranched poly(amido amine)s with tertiary amino cores for gene delivery.
Biomacromolecules
PUBLISHED: 05-30-2013
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Redox-responsive hyperbranched poly(amido amine)s (PAAs) with tertiary amino cores and amine, poly(ethylene glycol) (PEG) and hydroxyl terminal groups were prepared for DNA delivery respectively. The DNA condensation capability of PAAs was investigated using gel electrophoresis, and the results showed that PAA terminated with 1-(2-aminoethyl)piperazine (AEPZ) (BAA) is the most efficient in binding plasmid DNA (pDNA). The diameter and zeta-potential of polyplexes from PAAs were characterized using dynamic light scattering (DLS), and the morphology of the polyplexes was obtained using atomic force microscopy (AFM). All the PAAs were able to condense pDNA into nanoparticles with diameters between 50 and 200 nm with a positive surface charge when the weight ratio of polymer/DNA was higher than 20. Glutathione (GSH)-induced DNA release from polyplexes and the buffering capability of PAAs were investigated as well. Cytotoxicity of PAAs and in vitro gene transfection of polyplexes were evaluated in HEK293, COS-7, MCF-7 and Hep G2 cell lines, respectively. The results reflect that PAAs show remarkably low or even no cytotoxicity, and that PAA with amino terminal groups mediates the most efficient gene transfection with the transfection efficiency comparable to that of 25 kDa polyethylenimine. Further the effects of the presence of buthionine sulfoximine (BSO) on the transfection efficiency and cytotoxicity of BAA polyplexes were investigated.
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[Effects of non-saccharomyces albicans metabolic products on the proliferation of human umbilical vein endothelial cell ECV304].
Hua Xi Kou Qiang Yi Xue Za Zhi
PUBLISHED: 05-14-2013
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To evaluate the effects of non-Saccharomyces albicans metabolic products on the cell cycle distribution and proliferation of human umbilical vein endothelial cell ECV304 cells in vitro.
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Combined treatment with FK506 and nerve growth factor for spinal cord injury in rats.
Exp Ther Med
PUBLISHED: 05-04-2013
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Following spinal cord injury in rats, FK506 is able to protect local nerve tissue, promote neural regeneration, reduce neuronal apoptosis and accelerate the recovery of spinal cord functions. Nerve growth factor (NGF) is important in the regulation of central and peripheral nerve cell regeneration, growth differentiation and functions. Previous studies have shown that FK506 and NGF exhibit a synergistic effect in the treatment of peripheral nerve injury; however, it remains unclear whether the synergistic effect is present in the treatment of spinal cord injury. In this study, we combined FK506 and NGF for the treatment of spinal cord injury in rats. The NF200 protein expression in rats with spinal cord injury was determined using immunohistochemical staining and NF200 mRNA expression levels were observed using the reverse transcription-polymerase chain reaction method. The restoration of spinal cord functions was evaluated using the Basso, Beattie and Bresnahan score. The results demonstrated that the combined treatment significantly enhanced the expression of NF200 and improved spinal cord functions compared with the results of the single treatment. Our experimental observations indicated that FK506 and NGF exhibit a synergistic effect in the treatment of spinal cord injury in rats and that the combined treatment may effectively promote neural regeneration and functional recovery in rats following spinal cord injury.
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Genome-wide microRNA and messenger RNA profiling in rodent liver development implicates mir302b and mir20a in repressing transforming growth factor-beta signaling.
Hepatology
PUBLISHED: 04-26-2013
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MicroRNAs (miRNAs) are recently discovered small RNA molecules that regulate developmental processes, such as proliferation, differentiation, and apoptosis; however, the identity of miRNAs and their functions during liver development are largely unknown. Here we investigated the miRNA and gene expression profiles for embryonic day (E)8.5 endoderm, E14.5 Dlk1(+) liver cells (hepatoblasts), and adult liver by employing Illumina sequencing. We found that miRNAs were abundantly expressed at all three stages. Using K-means clustering analysis, 13 miRNA clusters with distinct temporal expression patterns were identified. mir302b, an endoderm-enriched miRNA, was identified as an miRNA whose predicted targets are expressed highly in E14.5 hepatoblasts but low in the endoderm. We validated the expression of mir302b in the endoderm by whole-mount in situ hybridization. Interestingly, mir20a, the most highly expressed miRNA in the endoderm library, was also predicted to regulate some of the same targets as mir302b. We found that through targeting Tgfbr2, mir302b and mir20a are able to regulate transforming growth factor beta (TGF?) signal transduction. Moreover, mir302b can repress liver markers in an embryonic stem cell differentiation model. Collectively, we uncovered dynamic patterns of individual miRNAs during liver development, as well as miRNA networks that could be essential for the specification and differentiation of liver progenitors. (HEPATOLOGY 2013).
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P58(IPK) inhibits coxsackievirus-induced apoptosis via the PI3K/Akt pathway requiring activation of ATF6a and subsequent upregulation of mitofusin 2.
Cell. Microbiol.
PUBLISHED: 04-25-2013
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Previously we found that prolonged endoplasmic reticulum (ER) stress caused by coxsackievirus B3 (CVB3) infection led to p58(IPK) downregulation and subsequent cell apoptosis. This finding implies that p58(IPK) expression benefits cell survival and counteracts CVB3-induced apoptosis. In testing this hypothesis, we first found that PI3K/Akt survival pathway is more sensitive than ERK1/2 in response to p58(IPK) expression. This finding was further verified by silencing p58(IPK) with specific siRNAs, which led to the significant suppression of phosphorylation of Akt (p-Akt) but not ERK1/2. Further, using CVB3-infected cell line expressing dominant negative ATF6a (DN-ATF6a), we found that expression of p58(IPK) and p-Akt was significantly reduced, which led to the decreased cell viability. However, when the DN-ATF6a cells were transiently transfected with p58(IPK) , an opposite result was obtained. Finally, by CVB3 infection of cells stably expressing p58(IPK) , we found that CVB3-induced mitochondria-mediated apoptosis was suppressed, which was evidenced by the reduced cytochrome c release and upregulation of the mitochondrial membrane protein mitofusin 2. However, silencing p58(IPK) with either specific siRNAs or DN-ATF6a sensitized cells to CVB3-induced apoptosis. These results suggest that p58(IPK) suppresses CVB3-induced apoptosis through selective activation of PI3K/Akt pathway that requires activation of ATF6a and subsequently upregulates mitofusin 2.
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An increased replication fidelity mutant of foot-and-mouth disease virus retains fitness in vitro and virulence in vivo.
Antiviral Res.
PUBLISHED: 03-22-2013
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In a screen for RNA mutagen-resistant foot-and-mouth disease virus (FMDV) strains, we isolated an FMDV mutant with RNA-dependent RNA polymerase (RdRp) R84H substitution. This mutant, selected under the mutagenic pressure of 5-fluorouracil (5-FU), is resistant not only to 5-FU but also to other two RNA mutagens, 5-azacytidine and ribavirin, suggesting that the RdRp R84H mutant is a high fidelity variant. Subsequently, the increased fidelity of this mutant was verified through analysis of mutation frequency, which revealed a 1.4-fold enhancement in RdRp fidelity compared with the wild-type virus. Further studies indicated that the R84H mutant exhibited slightly increased fitness in vitro, and its virulence was not reduced in suckling mice. These results indicated that an increase in RdRp fidelity does not always correlate with reduced virus fitness and virus attenuation. Thus, this isolated R84H mutant provides a new platform to examine the evolutionary dynamics of fidelity-changing RNA viruses, such as mutagen resistance, fitness and virulence.
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MiR-126 promotes coxsackievirus replication by mediating cross-talk of ERK1/2 and Wnt/?-catenin signal pathways.
Cell. Mol. Life Sci.
PUBLISHED: 03-19-2013
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Coxsackievirus B3 (CVB3) is one of the most prevalent causes of viral myocarditis and is associated with many other pathological conditions. CVB3 replication relies on host cellular machineries and causes direct damage to host cells. MicroRNAs have been found to regulate viral infections but their roles in CVB3 infection are still poorly understood. Here we describe a novel mechanism by which miR-126 regulates two signal pathways essential for CVB3 replication. We found that CVB3-induced ERK1/2 activation triggered the phosphorylation of ETS-1 and ETS-2 transcription factors, which induced miR-126 upregulation. By using both microRNA mimics and inhibitors, we proved that the upregulated miR-126 suppressed sprouty-related, EVH1 domain containing 1 (SPRED1) and in turn enhanced ERK1/2 activation. This positive feedback loop of ERK1/2-miR-126-ERK1/2 promoted CVB3 replication. Meanwhile, miR-126 expression stimulated GSK-3? activity and induced degradation of ?-catenin through suppressing LRP6 and WRCH1, two newly identified targets in the Wnt/?-catenin pathway, which sensitized the cells to virus-induced cell death and increased viral progeny release to initiate new infections. Our results demonstrate that upregulated miR-126 upon CVB3 infection targets SPRED1, LRP6, and WRCH1 genes, mediating cross-talk between ERK1/2 and Wnt/?-catenin pathways, and thus promoting viral replication and contributes to the viral cytopathogenicity.
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Desmoid tumor of posterior cruciate ligament of the knee: a case report.
BMC Musculoskelet Disord
PUBLISHED: 02-19-2013
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Desmoid tumor is a rare type of cancer that develops in the tissues that form tendons and ligaments. These tumors, also called aggressive fibromatosis, are considered benign with no metastatic potential. They may invade nearby tissues and organs, however, and can be difficult to control. Desmoid tumor in the posterior cruciate ligament (PCL) of the knee has never been described in the literature.
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Activity of antibacterial protein from maggots against Staphylococcus aureus in vitro and in vivo.
Int. J. Mol. Med.
PUBLISHED: 02-15-2013
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Maggots (larvae of Lucilia sericata) have shown therapeutic effects on refractory wounds infected with bacteria, yet the bacterial killing mechanisms are unclear. Herein, we report the isolation and purification of an antibacterial protein from maggots (MAMP). MAMP demonstrated inhibitory activity against both standard strains and clinically isolated antibiotic-resistant strains of Staphylococcus aureus in vitro. The topical use of MAMP effectively decreased the viability of S. aureus and promoted wound healing in an S. aureus mouse skin infection model. MAMP exerted its antibacterial activity via a bactericidal mechanism based on observations using scanning electron and transmission electron microscopy. MAMP interacted with the bacterial cell membrane and disrupted the cell surface structure. In addition, MAMP had weak hemolytic activity at a high concentration. Taken together, MAMP exhibits potential use as a topical agent for treating bacterial infections.
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An ERK-p38 subnetwork coordinates host cell apoptosis and necrosis during coxsackievirus B3 infection.
Cell Host Microbe
PUBLISHED: 01-16-2013
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The host response to a virus is determined by intracellular signaling pathways that are modified during infection. These pathways converge as networks and produce interdependent phenotypes, making it difficult to link virus-induced signals and responses at a systems level. Coxsackievirus B3 (CVB3) infection induces death of cardiomyocytes, causing tissue damage and virus dissemination, through incompletely characterized host cell signaling networks. We built a statistical model that quantitatively predicts cardiomyocyte responses from time-dependent measurements of phosphorylation events modified by CVB3. Model analysis revealed that CVB3-stimulated cytotoxicity involves tight coupling between the host ERK and p38 MAPK pathways, which are generally thought to control distinct cellular responses. The kinase ERK5 requires p38 kinase activity and inhibits apoptosis caused by CVB3 infection. By contrast, p38 indirectly promotes apoptosis via ERK1/2 inhibition but directly causes CVB3-induced necrosis. Thus, the cellular events governing pathogenesis are revealed when virus-host programs are monitored systematically and deconvolved mathematically.
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Butein inhibits NF-?B activation and reduces infiltration of inflammatory cells and apoptosis after spinal cord injury in rats.
Neurosci. Lett.
PUBLISHED: 01-08-2013
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The IKK/NF-?B signalling pathway plays a predominant role in the regulation of inflammation and apoptosis in spinal cord injury (SCI). We have previously demonstrated that targeting of the IKK/NF-?B pathway improved the recovery of locomotor function by reducing the infiltration of inflammatory cells and apoptosis after SCI in rats. Recently, the neuroprotective effects of butein have been shown via direct inhibition of the IKK/NF-?B pathway in vitro. In this study, we investigated the effects of butein on the IKK/NF-?B pathway in rats after SCI. Our results indicated that butein reduced the expression of NF-?B and activation of its inhibitor I-?B? at 24h after injury. Treatment with butein also resulted in significant inhibition of caspase-3 activation and neutrophil infiltration. Overall, our findings demonstrated the neuroprotective effects of butein in SCI in vivo and its potential mechanism. Our results further indicated that targeting of the IKK/NF-?B pathway may be useful in SCI therapy.
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Tissue-Specific and Cation/Anion-Specific DNA Methylation Variations Occurred in C. virgata in Response to Salinity Stress.
PLoS ONE
PUBLISHED: 01-01-2013
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Salinity is a widespread environmental problem limiting productivity and growth of plants. Halophytes which can adapt and resist certain salt stress have various mechanisms to defend the higher salinity and alkalinity, and epigenetic mechanisms especially DNA methylation may play important roles in plant adaptability and plasticity. In this study, we aimed to investigate the different influences of various single salts (NaCl, Na2SO4, NaHCO3, Na2CO3) and their mixed salts on halophyte Chloris. virgata from the DNA methylation prospective, and discover the underlying relationships between specific DNA methylation variations and specific cations/anions through the methylation-sensitive amplification polymorphism analysis. The results showed that the effects on DNA methylation variations of single salts were ranked as follows: Na2CO3> NaHCO3> Na2SO4> NaCl, and their mixed salts exerted tissue-specific effects on C. virgata seedlings. Eight types of DNA methylation variations were detected and defined in C. virgata according to the specific cations/anions existed in stressful solutions; in addition, mix-specific and higher pH-specific bands were the main type in leaves and roots independently. These findings suggested that mixed salts were not the simple combination of single salts. Furthermore, not only single salts but also mixed salts showed tissue-specific and cations/anions-specific DNA methylation variations.
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Organic carbon storage in Chinas urban areas.
PLoS ONE
PUBLISHED: 01-01-2013
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China has been experiencing rapid urbanization in parallel with its economic boom over the past three decades. To date, the organic carbon storage in Chinas urban areas has not been quantified. Here, using data compiled from literature review and statistical yearbooks, we estimated that total carbon storage in Chinas urban areas was 577 ± 60 Tg C (1 Tg = 10(12) g) in 2006. Soil was the largest contributor to total carbon storage (56%), followed by buildings (36%), and vegetation (7%), while carbon storage in humans was relatively small (1%). The carbon density in Chinas urban areas was 17.1 ± 1.8 kg C m(-2), about two times the national average of all lands. The most sensitive variable in estimating urban carbon storage was urban area. Examining urban carbon storages over a wide range of spatial extents in China and in the United States, we found a strong linear relationship between total urban carbon storage and total urban area, with a specific urban carbon storage of 16 Tg C for every 1,000 km(2) urban area. This value might be useful for estimating urban carbon storage at regional to global scales. Our results also showed that the fraction of carbon storage in urban green spaces was still much lower in China relative to western countries, suggesting a great potential to mitigate climate change through urban greening and green spaces management in China.
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[Effectiveness of a double-tsuge suture method in repairing Achilles tendon ruptures].
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi
PUBLISHED: 11-11-2011
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To investigate the effectiveness of a double-tsuge suture method with absorbable polydioxanone-cord (PDS-II) in repair of Achilles tendon ruptures.
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The immunity-related GTPase Irgm3 relieves endoplasmic reticulum stress response during coxsackievirus B3 infection via a PI3K/Akt dependent pathway.
Cell. Microbiol.
PUBLISHED: 11-10-2011
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The IRG protein Irgm3 preserves cell survival during coxsackievirus B3 (CVB3) infection. However, the molecular mechanisms are not clear. Here, we examined the effect of Irgm3 expression on ER stress triggered by pharmacological agents or CVB3 infection. In Tet-On/Irgm3 HeLa cells, Irgm3 expression suppressed either chemical- or CVB3-induced upregulation of glucose-regulated protein 78. Further, Irgm3 strongly inhibited the activation of both the PERK and ATF6 pathways of ER stress responses, which further led to the diminished phosphorylation of eIF2?, reduced cleavage/activation of transcription factor SREBP1 and attenuated induction of proapoptotic genes CHOP and GADD34. These data were further supported by experiments using Irgm3 knockout mouse embryonic fibroblasts, in which the ER stress induced by CVB3 was not relieved due to the lack of Irgm3 expression. In addition, the tunicamycin-triggered ER stress promoted the subsequent CVB3 infection. The effect of Irgm3 on ER stress and CVB3 infection was diminished by the PI3K inhibitor, LY294002, while inhibitors of ERK, JNK and p38 had no effect. These data were further corroborated by transfection of cells with a dominant negative Akt. Taken together, these data suggest that Irgm3 relieves the ER stress response via a PI3K/Akt dependent mechanism, which contributes to host defence against CVB3 infection.
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Adaptively deformed mesh based interface method for elliptic equations with discontinuous coefficients.
J Comput Phys
PUBLISHED: 11-06-2011
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Mesh deformation methods are a versatile strategy for solving partial differential equations (PDEs) with a vast variety of practical applications. However, these methods break down for elliptic PDEs with discontinuous coefficients, namely, elliptic interface problems. For this class of problems, the additional interface jump conditions are required to maintain the well-posedness of the governing equation. Consequently, in order to achieve high accuracy and high order convergence, additional numerical algorithms are required to enforce the interface jump conditions in solving elliptic interface problems. The present work introduces an interface technique based adaptively deformed mesh strategy for resolving elliptic interface problems. We take the advantages of the high accuracy, flexibility and robustness of the matched interface and boundary (MIB) method to construct an adaptively deformed mesh based interface method for elliptic equations with discontinuous coefficients. The proposed method generates deformed meshes in the physical domain and solves the transformed governed equations in the computational domain, which maintains regular Cartesian meshes. The mesh deformation is realized by a mesh transformation PDE, which controls the mesh redistribution by a source term. The source term consists of a monitor function, which builds in mesh contraction rules. Both interface geometry based deformed meshes and solution gradient based deformed meshes are constructed to reduce the L(?) and L(2) errors in solving elliptic interface problems. The proposed adaptively deformed mesh based interface method is extensively validated by many numerical experiments. Numerical results indicate that the adaptively deformed mesh based interface method outperforms the original MIB method for dealing with elliptic interface problems.
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PPE38 modulates the innate immune response and is required for Mycobacterium marinum virulence.
Infect. Immun.
PUBLISHED: 10-28-2011
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The proline-glutamic acid (PE) and proline-proline-glutamic acid (PPE) family proteins are prevalent in pathogenic mycobacteria and play a diverse role in mycobacterial pathogenesis. While some members have been studied, the function of most PE/PPE proteins remains unknown. In this study, we isolated a transposon-inactivated PPE38 mutant of Mycobacterium marinum and characterized its phenotype. We found that the PPE38 protein is associated with the cell wall and exposed on the cell surface. The inactivation of PPE38 altered the bacterial cell surface properties and led to deficiencies in cord formation, sliding motility, and biofilm formation. The PPE38 mutant was defective in phagocytosis by macrophages and exhibited reduced virulence in adult zebrafish. We also found that PPE38 is involved in the induction of proinflammatory cytokines in infected macrophages. Together, our results indicate that PPE38, a previously uncharacterized protein, plays a role in mycobacterial virulence, presumably by modulating the host innate immune response.
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Cu(2+)-selective fluorescent chemosensor based on coumarin and its application in bioimaging.
Dalton Trans
PUBLISHED: 09-14-2011
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A new fluorescent sensor L1 based on coumarin was synthesized. It shows high sensitivity and selectivity toward Cu(2+) in aqueous solution. The complexation mode and corresponding quenching mechanism were elucidated by ESI-MS and DFT calculations. In addition, biological imaging studies have demonstrated that L1 can detect Cu(2+) in living cells.
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The eukaryotic-type serine/threonine protein kinase Stk is required for biofilm formation and virulence in Staphylococcus epidermidis.
PLoS ONE
PUBLISHED: 05-26-2011
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Serine/threonine kinases are involved in gene regulation and signal transduction in prokaryotes and eukaryotes. Here, we investigated the role of the serine/threonine kinase Stk in the opportunistic pathogen Staphylococcus epidermidis.
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Controlled Delivery of Gentamicin Using Poly(3-hydroxybutyrate) Microspheres.
Int J Mol Sci
PUBLISHED: 05-23-2011
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Poly(3-hydroxybutyrate), P(3HB), produced from Bacillus cereus SPV using a simple glucose feeding strategy was used to fabricate P(3HB) microspheres using a solid-in-oil-water (s/o/w) technique. For this study, several parameters such as polymer concentration, surfactant and stirring rates were varied in order to determine their effect on microsphere characteristics. The average size of the microspheres was in the range of 2 ?m to 1.54 ?m with specific surface areas varying between 9.60 m(2)/g and 6.05 m(2)/g. Low stirring speed of 300 rpm produced slightly larger microspheres when compared to the smaller microspheres produced when the stirring velocity was increased to 800 rpm. The surface morphology of the microspheres after solvent evaporation appeared smooth when observed under SEM. Gentamicin was encapsulated within these P(3HB) microspheres and the release kinetics from the microspheres exhibiting the highest encapsulation efficiency, which was 48%, was investigated. The in vitro release of gentamicin was bimodal, an initial burst release was observed followed by a diffusion mediated sustained release. Biodegradable P(3HB) microspheres developed in this research has shown high potential to be used in various biomedical applications.
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A turn-on chemosensor for Hg2+ in aqueous media and its application in "MCT" imaging in living cells.
Dalton Trans
PUBLISHED: 05-16-2011
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A turn-on chemosensor L1, which exhibits high selectivity and sensitivity toward Hg(2+) over other common metal ions in aqueous media under a physiological pH window via a 1:1 binding mode, had been synthesized and characterized. L1 provides good fluorescent imaging of Hg(2+) in living cells. Particularly, we adopted the "micro computed tomography (MCT)" technology, successfully demonstrating the method of Hg(2+) sensing by L1 in cell lines, also the cell permeability of L1 and its imaging position in the cells.
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In vitro evaluation of 45S5 Bioglass®-derived glass-ceramic scaffolds coated with carbon nanotubes.
J Biomed Mater Res A
PUBLISHED: 05-05-2011
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Highly porous (> 90% porosity) 45S5 Bioglass®-derived glass-ceramic scaffolds were fabricated by foam replication method, and coated with carbon nanotubes (CNT) (coating thickness: 1 ?m) using electrophoretic deposition (EPD). In vitro cell culture using mesenchymal stem cells (MSCs) was carried out on both scaffold systems (with and without CNT coating) over a 4-week period. By using AlamarBlue™, BSA and alkaline phosphatase assays; the cell viability and differentiation were measured quantitatively measured and compared between the two scaffold types. The results showed that both scaffold systems are biocompatible with MSCs and they can support the cellular activity. No cytotoxic effects of CNT were observed under the conditions of the present experiments. Although a lower initial cell viability on the CNT-coated scaffolds was observed, no significant differences were found after 4 weeks of culture compared with the uncoated scaffolds. This work therefore shows that there is in principle no significant improvement of cellular responses by creating a CNT-coating on this type of highly bioactive scaffolds. However, the electrical conductivity introduced by the coating might have the potential to increase cell viability and differentiation when cell culture is carried out under the effect of electrical stimulation.
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[Short-term effectiveness of arthroscopic anterior cruciate ligament reconstruction with RetroButton-allogeneic tendon-interference screw].
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi
PUBLISHED: 04-20-2011
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To investigate the method and short-term effectiveness of arthroscopic reconstruction of anterior cruciate ligament (ACL) using RetroButton-allogeneic tendon-interference screw.
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Biocompatible hydroxyapatite nanoparticles as a redox luminescence switch.
J. Biol. Inorg. Chem.
PUBLISHED: 04-05-2011
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A redox luminescence switch was prepared by doping hydroxyapatite nanoparticles with CePO(4):Tb. The resulting multifunctional material exhibits good biocompatibility, biological affinity, and potential drug-carrying capability. The luminescent hydroxyapatite nanoparticles may find important applications in biomedical diagnostics, drug delivery, and biological sensors.
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Exposure to 3-methyl-4-nitrophenol affects testicular morphology and induces spermatogenic cell apoptosis in immature male rats.
Res. Vet. Sci.
PUBLISHED: 03-26-2011
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The pollutant 3-methyl-4-nitrophenol (PNMC), a major component of diesel exhaust particles, can cause many adverse health problems. In the present study, we investigated the effects of PNMC on the testes of Sprague Dawley rats treated subcutaneously for 5 days with different doses of PNMC (1, 10 or 100mg/kg). Exposure to PNMC caused a significant decrease in the plasma testosterone levels and in the absolute and relative weights of the testes. Severe histological lesions were observed in the testes of PNMC-treated animals. The ratio of the epithelial height to the seminiferous tubule diameter increased markedly in the PNMC-treated rats compared with the corresponding controls. In addition, PNMC exposure significantly increased the number of apoptotic spermatogenic cells compared with the controls whereas a significant decrease in the ratios of Bcl-2/Bax was observed at the same doses. These results suggest that PNMC exerts its gonadotoxicity in the rat mainly via apoptosis.
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Targeted delivery of mutant tolerant anti-coxsackievirus artificial microRNAs using folate conjugated bacteriophage Phi29 pRNA.
PLoS ONE
PUBLISHED: 03-18-2011
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Myocarditis is the major heart disease in infants and young adults. It is very commonly caused by coxsackievirus B3 (CVB3) infection; however, no specific treatment or vaccine is available at present. RNA interference (RNAi)-based anti-viral therapy has shown potential to inhibit viral replication, but this strategy faces two major challenges; viral mutational escape from drug suppression and targeted delivery of the reagents to specific cell populations.
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Identification of monochloro-nonabromodiphenyl ethers in the air and soil samples from south China.
Environ. Sci. Technol.
PUBLISHED: 03-14-2011
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Several studies have indicated that mixed brominated/chlorinated organic compounds could be formed during the thermal process such as the incineration of municipal solid waste and open burning of unregulated e-waste at recycling areas. In this study, air particles and soils from e-waste recycling areas, as well as outdoor and indoor air particles from urban Guangzhou, were collected and pooled for the identification of mixed chlorinated/brominated diphenyl ethers (PXDEs). Three monochloro-nonabromo diphenyl ethers (Cl-nonaBDEs), including 6-Cl-BDE-206, 5-Cl-BDE-207, and/or 4-Cl-BDE-208, were first structurally identified in these air and soil samples. The identification was done by comparison of retention times in chromatograms of pure reference compounds and environmental samples, as well as by comparison with full-scan mass spectra data in electron capture negative ionization mode. Because of their similar physical-chemical properties, 4-Cl-BDE-208 and 5-Cl-BDE-207 absolutely coeluted, even on a nonpolar DB-5 column. Further investigation is still needed to clarify these findings. Nevertheless, the results indicated that Cl-nonaBDEs would occur in various environmental matrices. Because the replacement of Br by Cl will change the physical-chemical properties of PBDE analogues, environmental occurrence, fate, and transport, the potential toxicity of PXDEs should be investigated.
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A rhodamine-based "turn-on" fluorescent chemodosimeter for Cu2+ and its application in living cell imaging.
J. Inorg. Biochem.
PUBLISHED: 02-17-2011
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A new fluorescent probe 1, N-(Rhodamine-6G)lactam-hydrazinecarbothioamide, was synthesized as a fluorescent and colorimetric chemodosimeter in aqueous solution for Cu(2+). Following Cu(2+)-promoted ring opening, redox and hydrolysis reactions, comparable amplifications of absorption and fluorescence signals were observed upon addition of Cu(2+); this suggests that chemodosimeter 1 effectively avoided the fluorescence quenching caused by the paramagnetic nature of Cu(2+). Importantly, 1 can selectively recognize Cu(2+) in aqueous media in the presence of other trace metal ions in organisms, abundant cellular cations and the prevalent toxic metal ions in the environment with high sensitivity (detection limit <3 ppb) and a rapid response time (<2 min). In addition, the biological imaging study has demonstrated that 1 can detect Cu(2+) in the living cells.
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Transition metal complexes of 2, 6-di ((phenazonyl-4-imino) methyl)-4-methylphenol: structure and biological evaluation.
Eur J Med Chem
PUBLISHED: 02-08-2011
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A symmetric ligand 2, 6-di ((phenazonyl-4-imino)methyl)-4-methylphenol (Dpmp) and its cobalt dinuclear complex (Co2(Dpmp)2(NO3)2(H2O)2·NO3·EtOH, (1) and zinc mononuclear complex Zn(Dpmp)(NO3)2, (2) have been prepared. The crystal structures were determined by single-crystal X-ray diffraction. The biological activity has been evaluated by examining their anti-oxidative activity and ability to bind to bovine serum albumin (BSA) and calf-thymus DNA (CT DNA) with UV-vis absorption, fluorescence, viscosity measurements and circular dichroism (CD) spectroscopies. The complexes exhibit good binding propensity to BSA and CT DNA. Both 1 and 2 have been found to promote cleavage of pUC19 DNA in the absence of any reducing agent. Antioxidant tests in vitro show the compounds possess significant antioxidant activity against superoxide and hydroxyl radicals.
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ygs is a novel gene that influences biofilm formation and the general stress response of Staphylococcus epidermidis.
Infect. Immun.
PUBLISHED: 12-20-2010
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Infections caused by the nosocomial pathogen Staphylococcus epidermidis frequently develop on implanted medical devices and involve biofilm formation. Biofilms are surface-attached microbial communities that show increased resistance to drug treatment and mechanisms of innate host defense. In this study, a mutant library of the clinical isolate S. epidermidis 1457 was constructed using mariner-based transposon mutagenesis. About a thousand mutants were screened, and 12 mutants were identified as significantly defective in biofilm formation. We focused on a mutant in which the transposon had inserted in a gene with unknown function, SERP0541, which is annotated as a gene encoding a GSP13-like general stress response protein. The gene was named ygs (encoding an unknown general stress protein). Various stresses, including heat, pH, high osmolarity, and ethanol affected the survival of the ygs mutant to a significantly higher degree than the wild-type strain and led to increased expression of ygs. Furthermore, synthesis of polysaccharide intercellular adhesin (PIA) and transcription of the PIA biosynthetic operon were significantly decreased in the ygs mutant. These results are in accordance with the putative involvement of ygs in stress-response gene regulation and indicate that ygs influences biofilm development by controlling PIA-dependent biofilm accumulation. Moreover, ygs had a significant impact on the formation of biofilms and metastatic disease in two catheter-related rat infection models. Our study shows that the ygs gene controls S. epidermidis biofilm accumulation and stress resistance, representing a key regulator of both structural and physiological biofilm characteristics with a significant impact on biofilm-associated infection.
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